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Results for “Biotechnology & applied microbiology”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 55 records · Page 3

Engineering the cellulolytic extreme thermophile Caldicellulosiruptor bescii to reduce carboxylic acids to alcohols using plant biomass as the energy source

Abstract Caldicellulosiruptor bescii is the most thermophilic cellulolytic organism yet identified (Topt 78 °C). It grows on untreated plant biomass and has an established genetic system thereby making it a promising microbial platform for lignocellulose conversion to bio-products. Here, we investigated the ability of engineered C. bescii to generate alcohols from carboxylic acids. Expression of aldehyde ferredoxin oxidoreductase (aor from Pyrococcus furiosus) and alcohol dehydrogenase (adhA from Thermoanaerobacter sp. X514) enabled C. bescii to generate ethanol from crystalline cellulose and from biomass by reducing the acetate produced by fermentation. Deletion of lactate dehydrogenase in a strain expressing the AOR–Adh pathway increased ethanol production. Engineered strains also converted exogenously supplied organic acids (isobutyrate and n-caproate) to the corresponding alcohol (isobutanol and hexanol) using both crystalline cellulose and switchgrass as sources of reductant for alcohol production. This is the first instance of an acid to alcohol conversion pathway in a cellulolytic microbe.

Biotechnology & Applied Microbiology↗

Bioremediation of heavy oily sludge: a microcosms study

Oily sludge is a residue from the petroleum industry composed of a mixture of sand, water, metals, and high content of hydrocarbons (HCs). The heavy oily sludge used in this study originated from Colombian crude oil with high density and low American Petroleum Institute (API) gravity. The residual waste from heavy oil processing was subject to thermal and centrifugal extraction, resulting in heavy oily sludge with very high density and viscosity. Biodegradation of the total petroleum hydrocarbons (TPH) was tested in microcosms using several bioremediation approaches, including: biostimulation with bulking agents and nutrients, the surfactant Tween 80, and bioaugmentation. Select HC degrading bacteria were isolated based on their ability to grow and produce clear zones on different HCs. Degradation of TPH in the microcosms was monitored gravimetrically and with gas chromatography (GC). The TPH removal in all treatments ranged between 2 and 67%, regardless of the addition of microbial consortiums, amendments, or surfactants within the tested parameters. The results of this study demonstrated that bioremediation of heavy oily sludge presents greater challenges to achieve regulatory requirements. Additional physicochemical treatments analysis to remediate this recalcitrant material may be required to achieve a desirable degradation rate.

59 BASIC BIOLOGICAL SCIENCES↗

Skinny kelp (Saccharina angustissima) provides valuable genetics for the biomass improvement of farmed sugar kelp (Saccharina latissima)

Abstract Saccharina latissima (sugar kelp) is one of the most widely cultivated brown marine macroalgae species in the North Atlantic and the eastern North Pacific Oceans. To meet the expanding demands of the sugar kelp mariculture industry, selecting and breeding sugar kelp that is best suited to offshore farm environments is becoming necessary. To that end, a multi-year, multi-institutional breeding program was established by the U.S. Department of Energy's (DOE) Advanced Research Projects Agency-Energy (ARPA-E) Macroalgae Research Inspiring Novel Energy Resources (MARINER) program. Hybrid sporophytes were generated using 203 unique gametophyte cultures derived from wild-collected Saccharina spp . for two seasons of farm trials (2019–2020 and 2020–2021). The wild sporophytes were collected from 10 different locations within the Gulf of Maine (USA) region, including both sugar kelp ( Saccharina latissima ) and the skinny kelp species ( Saccharina angustissima ). We harvested 232 common farm plots during these two seasons with available data. We found that farmed kelp plots with skinny kelp as parents had an average increased yield over the mean (wet weight 2.48 ± 0.90 kg m −1 and dry weight 0.32 ± 0.10 kg m −1 ) in both growing seasons. We also found that blade length positively correlated with biomass in skinny kelp x sugar kelp crosses or pure sugar kelp crosses. The skinny x sugar progenies had significantly longer and narrower blades than the pure sugar kelp progenies in both seasons. Overall, these findings suggest that sugar x skinny kelp crosses provide improved yield compared to pure sugar kelp crosses.

59 BASIC BIOLOGICAL SCIENCES↗

Global interfertility and heterosis in sugar kelp populations: a next step in sugar kelp breeding

Abstract The potential of seaweed aquaculture is restricted by high labor, production and processing costs, leading to low economic viability. Selective breeding can improve yields and cultivation efficiency, thereby decreasing production costs. Until now, genetic resources as input for Saccharina latissimabreeding trials have been sourced strictly locally, due to concerns regarding outplanting genetically exogenous material in local waters. Here we study, for the first time, worldwide interregional fertility of the seaweedS. latissima,in order to assess the potential of including globalS. latissimagenetic resources for selective breeding with regard to heterosis. We quantified the yield (as an indicative aquacultural performance) and morphological traits of intra- and interregionalS. latissimahybrids originating from a broad range of locations in a common garden experiment. Our results show that the practical application of worldwideS. latissimagenetic resources in breeding programs is feasible based on global interfertility. We found a wide morphological diversity of hybrids and observed significant heterosis in interregional hybrids. The degree of heterosis could not be linked to geographic distance. These findings reveal that worldwide genetic resources can considerably contribute toS. latissimabreeding programs and could offer a major next step in improving yields and quality traits.

Biotechnology & Applied Microbiology↗

Techno-economic and Life Cycle Analysis of MixAlco® Processes for Mixed Alcohol Production from Brown Algae

The need for producing renewable fuels from biomass has increased due to depleting fossil resources and environmental concerns. However, the low fraction of biomass carbon converted to product is an undeniable drawback for most current biofuel productions from fermentation due to undecomposed lignin in biomass composition and carbon loss as CO 2 . In this work, two main production routes of the MixAlco® process, the ketonization route (KR) and esterification route (ER) are evaluated for the mixed alcohol production by brown algae, a third-generation biomass without lignin. A novel fermentation process using syntrophic bacteria consortia (SBC) is developed to produce acetic acid from waste gas produced by KR and ER process. The paper investigates the integrated flowsheet for these alternative routes, using techno-economic and life cycle analysis to compare the minimum selling price and environmental impacts. From TEA, we find that the overall costs for KR and ER are lower than the SBC processes. The cost of ketonization routes is lower than esterification routes. The capital cost and operating cost for the ER+SBC process are the highest. Raw materials and utilities are the two major costs for all the processing routes examined. Here, the MSP for the ER+SBC process is the lowest out of all four routes. ER process performs the best in terms of environmental impacts except in water depletion compared with other processes, while the KR process performs the worst regarding the environmental metrics.

59 BASIC BIOLOGICAL SCIENCES↗

Ultrastructure of the green alga Dunaliella salina strain CCAP19/18 (Chlorophyta) as investigated by quick-freeze deep-etch electron microscopy

The single-celled green alga Dunaliella salina is a model system for studies on stress biology, in particular regarding secondary carotenoid accumulation. Under non-stress conditions the cells are green, but under abiotic stress the cells turn orange, because they switch their metabolism and accumulate β-Carotene in globules in the chloroplast. For the first time, Quick-freeze deep-etch electron microscopy was used to visualize cellular structures in green and orange cells of D. salina strain CCAP19/18. This allowed us to present an in-depth analysis of the cellular ultrastructure describing and comparing the features of the two cell types. Our images illustrate the presence of a pericellular matrix for this strain of D. salina. The pericellular matrix was spongy and strands of unknown material anchored it into the plasma membrane. The cytoplasm contained a variety of vesicles, vacuoles, and acidocalcisomes. We could show for strain CCAP19/18 that cytoplasmic lipid bodies were often in close proximity to and sometimes in contact with the outer chloroplast envelope membrane and with the endoplasmic reticulum. Major visible differences between green and orange cells were in the chloroplast: the orange cells have greatly reduced amounts of thylakoid membranes and greatly increased numbers of β-Carotene globules. We showed that the β-Carotene globules often made point contacts with thylakoid membranes, and frequently laid side-by-side along the thylakoid membrane surface, providing support to studies that indicated exchange of molecules between β-Carotene globules and thylakoid membranes. A unique finding was the β-Carotene globule duplets, suggesting intermediate stages in β-Carotene globule morphogenesis. Overall, the β-Carotene globules appear to be similar to plant plastoglobuli. We provide this description of the cellular ultrastructure features as a resource in the context of the recent publication of the genome of D. salina strain CCAP19/18 to expand on the knowledge regarding this novel reference strain.

59 BASIC BIOLOGICAL SCIENCES↗

Enhanced carbon-transfer and -utilization efficiencies achieved using membrane carbonation with gas sources having a range of CO 2 concentrations

The economic viability of microalgal biofuels relies on increasing productivity in a cost-effective manner. As microalgal biomass contains > 50% carbon, a high rate of CO 2 delivery is required for high productivity, and inefficient CO 2 delivery amplifies operating costs. Membrane carbonation using non-porous hollow fiber membranes can ideally deliver CO 2 without bubble formation and high carbon transfer efficiency. Because CO 2 streams from industrial resources are not 100% CO 2 , the buildup of inert gasses can significantly lower the CO 2 delivery rate when the distal end of the membrane is closed. To overcome the buildup of inert gases, we managed the distal end of the membranes with three different approaches: fully open end, restricted bleed valve, and restricted bleed valve with pH-actuated venting. For all approaches, CO 2 was delivered to membranes ondemand based on a pH set point. Evaluating a wide range of CO 2 concentrations (10% to 100%), we found that all approaches eliminated the buildup of inert gases, could maintain target pH values and gave the same biomass productivities and carbon distributions. However, carbon transfer efficiency depended on the operation of the distal end. Fully open-end operation gave a poor carbon transfer efficiency because of excessive loss of CO 2 from the distal end. However, restricting the exit flow rate to ≤4 cm 3 /min mitigated the problems of excessive CO 2 loss, but without incurring a large loss of CO 2 -delivery flux. For the continuous cultivation, combining a restricted bleed valve with pH-actuated venting improved the carbon-transfer efficiency and -utilization efficiencies up to 85% and 67%, respectively, with a sufficient CO 2 delivery flux.

42 ENGINEERING↗

A long-term pilot-scale algal cultivation on power plant flue gas – Cultivation stability and biomass accumulation

A long-term culture of a selected freshwater microalgal assemblage on power plant flue gas was realized using a pilot-scale photobioreactor. Different culture conditions of light cycle, harvesting amount, pH stabilization, and sulfur addition were tested during a 12-month continuous operation at a natural gas fired power plant. The results concluded that the daily harvesting amount of 60% with no light-cycle and NH 4 NO 3 as the nitrogen source and pH stabilization agent, resulted in optimal cultivation performance. Biomass concentration and biomass productivity reached 1.07 ± 0.23 g L -1 and 0.43 ± 0.14 g L -1 d -1 , respectively. During the year-long cultivation period, the algal assemblage was relatively stable with average abundances of 88 ± 4% and 12 ± 4% for algae and bacteria, respectively, and no significant differences between individual case scenarios. The stable culture fostered a stable biomass composition. Protein, carbohydrate, and lipid contents of the algal biomass were 58.60 ± 2.41%, 19.5 ± 2.44%, and 9.70 ± 1.68%, respectively.

09 BIOMASS FUELS↗

Adaptive Laboratory Evolution for algal strain improvement: methodologies and applications

Originally developed in bacterial and fungal models, Adaptive Laboratory Evolution (ALE) experimentally selects for targeted phenotypes by allowing cells containing beneficial random genetic mutations to replicate more quickly in the presence of environmental stress. When applied to microalgae, this technique has been used to increase growth rate, stress tolerance, substrate utilization, and product yield. This review discusses ALE experimental design parameters, and summarizes key results and lessons learned from recent microalgal ALE studies. By documenting the current state of the field, we provide insight on the ways microalgal ALE might be used to develop commercially viable biotechnology strains.

09 BIOMASS FUELS↗